Rapid three-dimensional imaging and analysis of the beating embryonic heart reveals functional changes during development
Rapid three-dimensional imaging and analysis of the beating embryonic heart reveals functional changes during development
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DOI：
10.1002/dvdy.20926
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发表时间：
2006-11-01
影响因子：
2.5
通讯作者：
Dickinson, Mary E.
中科院分区：
文献类型：
--
作者：
Liebling, Michael;Forouhar, Arian S.;Dickinson, Mary E.
我们报告了一种精确的方法来研究跳动的胚胎斑马鱼心脏的功能动力学。由于心脏的快速收缩和血细胞的高速运动，使得在体内研究与心脏功能相关的细胞和亚细胞事件变得困难。我们设计了一种动态三维采集、重建和分析程序，结合了（1）新开发的共聚焦狭缝扫描显微镜，（2）用于收集和同步循环图像序列的新策略，以在跳动心脏的整个深度上建立具有高时间和空间分辨率的体积，以及（3）用于系统提取定量信息以描述表型和功能的数据分析和简化方案。我们已经使用这种方法来表征血流和心脏效率的成像荧光蛋白表达的血液和内皮细胞的心脏从一个管发展到一个多腔器官。该方法是足够强大的图像组织内的心脏在细胞分辨率在很宽的年龄范围内，即使运动模式只是准周期性的。这些工具可推广到在微观尺度上成像和分析其他周期性运动的结构。
We report an accurate method for studying the functional dynamics of the beating embryonic zebrafish heart. The fast cardiac contraction rate and the high velocity of blood cells have made it difficult to study cellular and subcellular events relating to heart function in vivo. We have devised a dynamic three-dimensional acquisition, reconstruction, and analysis procedure by combining (1) a newly developed confocal slit-scanning microscope, (2) novel strategies for collecting and synchronizing cyclic image sequences to build volumes with high temporal and spatial resolution over the entire depth of the beating heart, and (3) data analysis and reduction protocols for the systematic extraction of quantitative information to describe phenotype and function. We have used this approach to characterize blood flow and heart efficiency by imaging fluorescent protein-expressing blood and endocardial cells as the heart develops from a tube to a multichambered organ. The methods are sufficiently robust to image tissues within the heart at cellular resolution over a wide range of ages, even when motion patterns are only quasiperiodic. These tools are generalizable to imaging and analyzing other cyclically moving structures at microscopic scales.